Purification and characterization of soluble human IL-6 receptor expressed in CHO cells.
An immunosorbent assay system to detect genetically engineered IL-6 receptor (IL-6R) was established, whereby soluble IL-6 receptor (sIL-6R) was detected in the culture medium when sIL-6R cDNA was transfected into COS1 cells. A stably transformed Chinese hamster ovary (CHO) cell line constitutively expressing sIL-6R has been established. The recombinant sIL-6R was purified to homogeneity by sequential filtration and chromatography of the culture medium. The recombinant sIL-6R augmented the sensitivity of M1 cells to IL-6 in growth inhibition assay in a dose-dependent manner. Furthermore, a radioisotope immunosorbent assay (RIA) utilizing recombinant sIL-6R was established which could detect IL-6 in a quantity as low as 10 ng/ml.
Yasukawa
K
K
Biotechnology Research Laboratory, Tosoh Corporation, Kanagawa.
Saito
T
T
Fukunaga
T
T
Sekimori
Y
Y
Koishihara
Y
Y
Fukui
H
H
Ohsugi
Y
Y
Matsuda
T
T
Yawata
H
H
Hirano
T
T
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D002455:Cell Division; D002460:Cell Line; D002851:Chromatography, High Pressure Liquid; D006224:Cricetinae; D004247:DNA / Q000737:chemistry; D006801:Humans; D008969:Molecular Sequence Data; D011971:Receptors, Immunologic / Q000096:biosynthesis / Q000235:genetics*; D019947:Receptors, Interleukin-6; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000235:genetics; D012995:Solubility
D011971:Receptors, Immunologic; D019947:Receptors, Interleukin-6; D011994:Recombinant Proteins; D004247:DNA